US20130072565A1 - Method of increasing cell proliferation by advanced wound dressings - Google Patents
Method of increasing cell proliferation by advanced wound dressings Download PDFInfo
- Publication number
- US20130072565A1 US20130072565A1 US13/623,564 US201213623564A US2013072565A1 US 20130072565 A1 US20130072565 A1 US 20130072565A1 US 201213623564 A US201213623564 A US 201213623564A US 2013072565 A1 US2013072565 A1 US 2013072565A1
- Authority
- US
- United States
- Prior art keywords
- dialkyl carbamoyl
- hydrophobic substance
- cell proliferation
- composition according
- carbamoyl chloride
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 238000000034 method Methods 0.000 title claims abstract description 15
- 230000004663 cell proliferation Effects 0.000 title abstract description 30
- 208000027418 Wounds and injury Diseases 0.000 claims abstract description 48
- 206010052428 Wound Diseases 0.000 claims abstract description 42
- 125000005117 dialkylcarbamoyl group Chemical group 0.000 claims description 45
- 230000002209 hydrophobic effect Effects 0.000 claims description 26
- 239000000126 substance Substances 0.000 claims description 24
- 210000004027 cell Anatomy 0.000 claims description 19
- 210000002950 fibroblast Anatomy 0.000 claims description 19
- 230000029663 wound healing Effects 0.000 claims description 18
- -1 alkyl ketene dimer Chemical compound 0.000 claims description 17
- 239000000203 mixture Substances 0.000 claims description 17
- 230000035876 healing Effects 0.000 claims description 10
- 230000035755 proliferation Effects 0.000 claims description 9
- 150000003839 salts Chemical class 0.000 claims description 6
- 230000002708 enhancing effect Effects 0.000 claims description 4
- CKDWPUIZGOQOOM-UHFFFAOYSA-N Carbamyl chloride Chemical compound NC(Cl)=O CKDWPUIZGOQOOM-UHFFFAOYSA-N 0.000 claims description 3
- 210000002510 keratinocyte Anatomy 0.000 claims description 3
- 150000002148 esters Chemical class 0.000 claims 4
- WHBHBVVOGNECLV-OBQKJFGGSA-N 11-deoxycortisol Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 WHBHBVVOGNECLV-OBQKJFGGSA-N 0.000 claims 1
- 239000003937 drug carrier Substances 0.000 claims 1
- 239000000546 pharmaceutical excipient Substances 0.000 claims 1
- 230000004936 stimulating effect Effects 0.000 abstract description 2
- 239000000463 material Substances 0.000 description 11
- 239000002609 medium Substances 0.000 description 10
- 230000006378 damage Effects 0.000 description 9
- 238000002835 absorbance Methods 0.000 description 6
- 238000011156 evaluation Methods 0.000 description 6
- 208000014674 injury Diseases 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 239000004744 fabric Substances 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 239000003102 growth factor Substances 0.000 description 4
- 230000035752 proliferative phase Effects 0.000 description 4
- 238000007634 remodeling Methods 0.000 description 4
- 210000001772 blood platelet Anatomy 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 239000010410 layer Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 210000003491 skin Anatomy 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 2
- 102000009123 Fibrin Human genes 0.000 description 2
- 108010073385 Fibrin Proteins 0.000 description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000001574 biopsy Methods 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 230000002500 effect on skin Effects 0.000 description 2
- 210000001339 epidermal cell Anatomy 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- 230000003328 fibroblastic effect Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000002356 single layer Substances 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- ZNQVEEAIQZEUHB-UHFFFAOYSA-N 2-ethoxyethanol Chemical compound CCOCCO ZNQVEEAIQZEUHB-UHFFFAOYSA-N 0.000 description 1
- 238000012935 Averaging Methods 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 208000034656 Contusions Diseases 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 239000006145 Eagle's minimal essential medium Substances 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 208000035874 Excoriation Diseases 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 102000016359 Fibronectins Human genes 0.000 description 1
- 108010067306 Fibronectins Proteins 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 238000012313 Kruskal-Wallis test Methods 0.000 description 1
- 208000034693 Laceration Diseases 0.000 description 1
- 238000000585 Mann–Whitney U test Methods 0.000 description 1
- BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 description 1
- 102000016611 Proteoglycans Human genes 0.000 description 1
- 108010067787 Proteoglycans Proteins 0.000 description 1
- 208000002847 Surgical Wound Diseases 0.000 description 1
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 1
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 238000002266 amputation Methods 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 230000002491 angiogenic effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 230000033077 cellular process Effects 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 230000037319 collagen production Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000009519 contusion Effects 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000037313 granulation tissue formation Effects 0.000 description 1
- KIUKXJAPPMFGSW-MNSSHETKSA-N hyaluronan Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)C1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H](C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-MNSSHETKSA-N 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229940099552 hyaluronan Drugs 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 238000010874 in vitro model Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- PQLXHQMOHUQAKB-UHFFFAOYSA-N miltefosine Chemical compound CCCCCCCCCCCCCCCCOP([O-])(=O)OCC[N+](C)(C)C PQLXHQMOHUQAKB-UHFFFAOYSA-N 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 230000002297 mitogenic effect Effects 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 208000019180 nutritional disease Diseases 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 229960003531 phenolsulfonphthalein Drugs 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000035409 positive regulation of cell proliferation Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000035807 sensation Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 210000005167 vascular cell Anatomy 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/16—Amides, e.g. hydroxamic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/28—Polysaccharides or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
Definitions
- the present invention relates generally to a method to stimulate cell proliferation in a wound. More specifically this invention relates to wound healing, due to this increased cell proliferation.
- wound healing is an intricate process in which the skin (or another organ) repairs itself after injury.
- Healing like all other biological processes, is a cellular process.
- the occurrence of a wound immediately triggers the onset of this process, which continues until the injury is healed.
- wound refers to tissue damage or loss of any kind, including but not limited to, cuts, incisions (including surgical incisions), abrasions, lacerations, contusions, bums, amputations and the like. Rapid healing of a wound reduces long term healthcare costs and improves patient recovery, including, regaining of sensation, function and aesthetics.
- hydrophobic substances e.g. dialkyl carbamoyl chloride or alkyl ketene dimer that preferably could be associated with a carrier to increase cell proliferation, which is ideal for treatment of wounds.
- This invention is based on the unexpected findings that hydrophobic substances including but not limited to dialkyl carbamoyl chloride and alkyl ketene dimer stimulate cell proliferation.
- the present invention discloses the use of hydrophobic substances to induce cell proliferation in wounds.
- hydrophobic substances inducing cell proliferation is ideal for treatment of wounds, especially wounds with delayed healing process due to reduced cell proliferation and wounds where a quick healing process is desirable.
- FIG. 2 shows the BWAT score on day 1, 5, 10 and 15 for patients treated with dialkyl carbamoyl chloride treated wound dressings for enhanced cell and for patient treated with untreated wound dressings.
- the present invention is ideal for treatment of certain wounds.
- this method of treatment stimulates cell proliferation thus enhancing wound healing.
- Wound healing is divided into three sequential, yet overlapping, phases: the inflammatory phase, the proliferative phase and the remodeling phase.
- the inflammatory phase The bleeding of the wound initiates thrombocyte aggregation at the injury site to form a coagel comprised of fibrin and thrombocytes, in order to stop the bleeding and also to protect the wound from bacterial contamination and fluid loss. Bacteria, dead tissue and foreign substances are phagocytosed and removed by macrophages. During this phase the formation of a provisional extracellular matrix take place, comprising fibrins, fibronectins and glycosaminoglycans. Inflammation proceeds until the wound is cleaned from unwanted substances and microorganisms.
- the proliferative phase Macrophages and thrombocytes are important factors in the proliferative phase as well due to the production and secretion of growth factors.
- the growth factors stimulate proliferation of endothelial cells, fibroblasts and smooth muscle cells. These cells will eventually replace the inflammatory cells in the wound.
- angiogenesis in this phase, new blood vessels are formed that supply the wound. with oxygen.
- Fibroblasts grow and initiate collagen production, which leads to granulation tissue formation resulting in reproduction of dermis and subcutis.
- the epithelium as well begins to reproduce by proliferation of keratinocytes.
- the wound is gradually made smaller.
- the remodeling phase During the remodeling phase the primitive extracellular matrix is degraded and simultaneously remodeling of a permanent matrix made up of collagen, elastin and proteoglycans take place.
- Some patients have reduced cell proliferation, due to e.g., chemotherapy, age or nutritional diseases. This reduced cell proliferation can cause impaired wound healing.
- the present invention stimulates cell proliferation and is therefore ideal for treatment of certain wounds, especially wounds with a delayed healing process, due to reduced cell proliferation, and wounds where a quick healing process is desirable.
- U.S. Pat. No. 6.699,287 discloses a dermal scaffold with wound healing effect by constituting microenvironments suitable for migration and proliferation of fibroblasts and vascular cells surrounding the wound.
- Patent application EP1637145 discloses an invention related to a wound healing promoting material. This material utilizes blood cells having wound healing effects which can improve the growth of fibroblasts, and allow them to produce growth factors involved in wound healing and thereby promoting wound healing.
- an invention is described that provides a composition capable of stimulating growth and regeneration of epidermal cells.
- the composition comprises an aqueous, cell-free extract derived from epidermal cells effective to inhibit fibroblast proliferation in the wound region.
- the principle of binding microorganisms with hydrophobic dressings discussed above is a modem and overall effective Method for anti-microbial wound healing, which is described in U.S. Pat. No. 4,617,326 and U.S. Patent application 2006/0129080.
- hydrophobic substances e.g., dialkyl carbamoyl chloride or alkyl ketene dimer
- a carrier to induce increased cell proliferation, including but not limited to the proliferation of fibroblasts and keratinocytes.
- hydrophobic substances as for example dialkyl carbamoyl chloride or alkyl ketene dimer
- the present invention may be used to enhance the natural healing process in wounds.
- the invention disclosed herein is based on the unexpected finding that hydrophobic substances promote cell proliferation.
- human dermal fibroblasts (CCL-110, ATCC/LGC-Standards AB, Boras, Sweden) were cultured in a culture incubator at 37° C. in 5% CO 2 atmosphere (Forma Science; AB Ninolab), using Eage's minimal essential medium with Earle's salts (EMEM, Sigma-Aldrich), supplemented with 10% foetal calf serum (FCS; Sigma-Aldrich). All handling of cells were carried out aseptically in a laminar airflow bench (Holten 2448; Ninolab).
- a sodium salt (XTT-assay, Sigma, USA) was used to study the influence on cell proliferation caused by the dialkyl carbamoyl chloride treated wound dressing as described in Example 5. Depending on the intra cellular dehydrogenase activity of the mitochondria, this salt will cause a colour change. The colour change is directly proportional to the increased number of cells and hence also directly proportional to cell proliferation. The colour change was measured using a 96-well plate reader (VMaz Kinetic ELISA; Molecular Devices) and software was used to calculate the absorbance at 450 nm (Softmax Pro, Molecular Devices). Cells were distributed to wells containing dialkyl carbamoyl chloride treated wound dressing material and culture medium and 12 wells only containing culture medium. Sterile disposable tools of 5 mm (Disposable Biopsy Punsch, Miltex, USA) were used to punch suitable pieces of the sterile wound dressing, which then were placed into the wells.
- Dialkyl Carbamoyl Chloride Increases Cell Proliferation In Vivo
- Twenty patients with wounds after transplantation of skin to another location are admitted to this open study.
- the patients are included in regularly planned follow ups with the medical staff. Inclusion criteria are the presence of a wound after skin has been removed for transplantation not more than 24 hours ago.
- the patients arc treated once daily using dressings manufactured according to example 3 herein.
- Ten of the admitted patients are treated with these cell proliferation inducing and ten patients serve as a control group treated with dressings without dialkyl carbamoyl chloride.
- BWAT Bates-Jensen wound assessment tool
- Fibroblasts cultured according to example 1 were grown until confluence at the start of the experiment.
- a mechanical injury was created by scraping the monolayer of cultured cells with a 10 ⁇ l sterile plastic pipette (D10ST, Gilson, INc.), forming a cross.
- the resulting damage was measured with a calibrated size marker at several time points, using a image processing software.
- cell proliferation and migration was identified by measuring the distance between the cells. To accomplish this, a line was drawn parallel to the wound edges, with multiple lines then drawn perpendicular to this, across the damaged area. This allowed a mean value for the length of the damaged area to be calculated. To reproduce measurements between incubation times, calculations were always made in the same the same area. Creating a cross in each culture well made it easy to identify this area by locating the centre of the cross and always beginning in the upper right arm of the cross.
- Healing rate was calculated by dividing the restored distance by the time between each measurement. Measurements were performed at 1, 3, 6, 24, 32, 48, 56 and 72 hours, after creating the mechanical damage. In the wound-healing model, the compresses were cut into small pieces using the 5 mm biopsy punch (used in example 2). A total of 12 wells was used, with six wells used for the dialkyl carbamoyl chloride treated wound dressings as described in example 5 and six wells with medium only. Care was taken not to contaminate the culture wells with dressing debris or fibres.
- the hydrophobic layer is preferably produced by applying to a cellulose acetate fabric an amount of dialkyl carbamoyl chloride, making a covalent bond between the materials.
- the dialkyl carbamoyl chloride is applied by letting the fabric pass through a bath of 0.5-2% carbamoyl chloride solution. The applied amount is checked by using the hydrophobicity test below.
- the acetate fabric is on rolls of 50 m length and at a width of 1 m, and taken as such to the next step.
- a fluid comprising formamide and ethylene glycol-monoethyl-ether with known dyne level between 37 and 50 dyne/cm.
- test sample of acetate woven is 0.1 m 2 .
- the result is documented for the batch in question.
- Results The sample shall have a dyne level less than 42 dyne/cm Cutting and adding of carrier layer.
- the now bonded hydrophobic layer is cut into suitable size pieces for the final product, in this case 30 mm ⁇ 30 mm.
- the cut pieces are placed on carrier-film, prepared with the adhesive and release paper and in strips of 100 mm.
- the bonded pieces are centered on the carrier strips and with a distance from center to center of 80 mm.
- a release paper of the same width is finally then applied on inner side.
- the now completely assembled dressing is cut into pieces of 80 mm ⁇ 100 mm, sterilized and then packed 10 by 10 in cartons.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Hematology (AREA)
- Materials Engineering (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dermatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Materials For Medical Uses (AREA)
Abstract
The present invention relates generally to a method for stimulating cell proliferation. More specifically this invention discloses the use of increased cell proliferation in treatment of certain wounds.
Description
- This application claims priority from U.S. Provisional Application Ser. No. 61/626,136 filed Sep. 21, 2011.
- The present invention relates generally to a method to stimulate cell proliferation in a wound. More specifically this invention relates to wound healing, due to this increased cell proliferation.
- Wound healing is an intricate process in which the skin (or another organ) repairs itself after injury. Healing, like all other biological processes, is a cellular process. The occurrence of a wound immediately triggers the onset of this process, which continues until the injury is healed. As used herein, the term “wound” refers to tissue damage or loss of any kind, including but not limited to, cuts, incisions (including surgical incisions), abrasions, lacerations, contusions, bums, amputations and the like. Rapid healing of a wound reduces long term healthcare costs and improves patient recovery, including, regaining of sensation, function and aesthetics.
- It is an object of the present invention to provide the use of hydrophobic substances, e.g. dialkyl carbamoyl chloride or alkyl ketene dimer that preferably could be associated with a carrier to increase cell proliferation, which is ideal for treatment of wounds. This invention is based on the unexpected findings that hydrophobic substances including but not limited to dialkyl carbamoyl chloride and alkyl ketene dimer stimulate cell proliferation.
- The present invention discloses the use of hydrophobic substances to induce cell proliferation in wounds.
- The use of hydrophobic substances inducing cell proliferation is ideal for treatment of wounds, especially wounds with delayed healing process due to reduced cell proliferation and wounds where a quick healing process is desirable.
- Other objects and features of the invention will be more fully apparent from the following disclosure and appended claims.
-
FIG. 1 shows the enhanced cell proliferation for fibroblasts associated with a carbamoyl chloride treated fabric compared to untreated control cells. (p=0,028: n=8), -
FIG. 2 shows the BWAT score onday -
FIG. 3 shows the restoration of damaged fibroblast surface. By 72 hours, the restoration differed significantly between the dialkyl carbamoyl chloride group and untreated medium control (p=0.04331; n=6). - The present invention is ideal for treatment of certain wounds. In a preferred embodiment, this method of treatment stimulates cell proliferation thus enhancing wound healing.
- Wound healing is divided into three sequential, yet overlapping, phases: the inflammatory phase, the proliferative phase and the remodeling phase.
- The inflammatory phase. The bleeding of the wound initiates thrombocyte aggregation at the injury site to form a coagel comprised of fibrin and thrombocytes, in order to stop the bleeding and also to protect the wound from bacterial contamination and fluid loss. Bacteria, dead tissue and foreign substances are phagocytosed and removed by macrophages. During this phase the formation of a provisional extracellular matrix take place, comprising fibrins, fibronectins and glycosaminoglycans. Inflammation proceeds until the wound is cleaned from unwanted substances and microorganisms.
- The proliferative phase. Macrophages and thrombocytes are important factors in the proliferative phase as well due to the production and secretion of growth factors. The growth factors stimulate proliferation of endothelial cells, fibroblasts and smooth muscle cells. These cells will eventually replace the inflammatory cells in the wound. In the angiogenesis in this phase, new blood vessels are formed that supply the wound. with oxygen. Fibroblasts grow and initiate collagen production, which leads to granulation tissue formation resulting in reproduction of dermis and subcutis. The epithelium as well begins to reproduce by proliferation of keratinocytes. During the proliferative phase the wound is gradually made smaller.
- The remodeling phase. During the remodeling phase the primitive extracellular matrix is degraded and simultaneously remodeling of a permanent matrix made up of collagen, elastin and proteoglycans take place.
- Some patients have reduced cell proliferation, due to e.g., chemotherapy, age or nutritional diseases. This reduced cell proliferation can cause impaired wound healing. The present invention stimulates cell proliferation and is therefore ideal for treatment of certain wounds, especially wounds with a delayed healing process, due to reduced cell proliferation, and wounds where a quick healing process is desirable.
- Further, there are several in inventions enhancing wound healing by stimulation of cell proliferation. However, none of the following inventions utilize hydrophobic substances. U.S. Pat. No. 6.699,287 discloses a dermal scaffold with wound healing effect by constituting microenvironments suitable for migration and proliferation of fibroblasts and vascular cells surrounding the wound.
- In U.S. Pat. No. 5,457,093, gel formulations are described containing polypeptide growth factors having human mitogenic or angiogenic activity used for wound healing.
- Patent application EP1637145 discloses an invention related to a wound healing promoting material. This material utilizes blood cells having wound healing effects which can improve the growth of fibroblasts, and allow them to produce growth factors involved in wound healing and thereby promoting wound healing.
- In U.S. Pat. No. 5,720,981, an invention is described that provides a composition capable of stimulating growth and regeneration of epidermal cells. The composition comprises an aqueous, cell-free extract derived from epidermal cells effective to inhibit fibroblast proliferation in the wound region. The principle of binding microorganisms with hydrophobic dressings discussed above is a modem and overall effective Method for anti-microbial wound healing, which is described in U.S. Pat. No. 4,617,326 and U.S. Patent application 2006/0129080.
- However, none of the prior art describes the use of hydrophobic substances, e.g., dialkyl carbamoyl chloride or alkyl ketene dimer, optionally applied to a carrier to induce increased cell proliferation, including but not limited to the proliferation of fibroblasts and keratinocytes.
- Surprisingly, we have discovered that hydrophobic substances, as for example dialkyl carbamoyl chloride or alkyl ketene dimer, stimulate cell proliferation, for example in wounds and may when applied to a carrier be used as wound dressings for improved wound healing. The present invention may be used to enhance the natural healing process in wounds. The invention disclosed herein is based on the unexpected finding that hydrophobic substances promote cell proliferation.
- Other objects and features of the invention will be more fully apparent from the following examples and appended claims.
- Commercially available human dermal fibroblasts (CCL-110, ATCC/LGC-Standards AB, Boras, Sweden) were cultured in a culture incubator at 37° C. in 5% CO2 atmosphere (Forma Science; AB Ninolab), using Eage's minimal essential medium with Earle's salts (EMEM, Sigma-Aldrich), supplemented with 10% foetal calf serum (FCS; Sigma-Aldrich). All handling of cells were carried out aseptically in a laminar airflow bench (Holten 2448; Ninolab).
- Fibroblasts were initiated in culture, as previously described by Falk P. Experimental Models of the Human Perioneal Environment: Effects of TGF-beta and Hyaluronan; University of Gothenburg, 2008.
- Prior to the start of the experiments performed in example 2 and 3, cells were cultured to approximately 50% confluence and then preincubated for 24 hours in a medium containing 1% FCS only. Cell count was done prior to the start of the experiments of example 2 and 3 to verify that here was no difference in cell count between the different wells.
- Throughout the examples 2 and 3 below the non-parametric Kruskal-Wallis test was used to detect overall differences. with the Mann Whitney U test performed between individual groups. A p-value <0.05 was considered statistically significant. All calculations were performed using StatView software (v5.0; Abacus Concepts).
- A sodium salt (XTT-assay, Sigma, USA) was used to study the influence on cell proliferation caused by the dialkyl carbamoyl chloride treated wound dressing as described in Example 5. Depending on the intra cellular dehydrogenase activity of the mitochondria, this salt will cause a colour change. The colour change is directly proportional to the increased number of cells and hence also directly proportional to cell proliferation. The colour change was measured using a 96-well plate reader (VMaz Kinetic ELISA; Molecular Devices) and software was used to calculate the absorbance at 450 nm (Softmax Pro, Molecular Devices). Cells were distributed to wells containing dialkyl carbamoyl chloride treated wound dressing material and culture medium and 12 wells only containing culture medium. Sterile disposable tools of 5 mm (Disposable Biopsy Punsch, Miltex, USA) were used to punch suitable pieces of the sterile wound dressing, which then were placed into the wells.
- During proliferation studies, a medium without phenol red wa used to reduce the red background signal in the XTT assay. Absorbance at 450 nm (absorbance maximum), 650 nm (background absorbance) was measured. A total of eight wells were used in each setup for proliferation: four wells containing fibroblasts in the presence of dialkyl carbamoyl chloride treated dressings placed in the medium, four plates serving as untreated control (the experiment was run twice).
- The results showed increased cell proliferation in wells containing the dialkyl carbamoyl chloride treated material compared to control wells. Absorbance in wells containing dialkyl carbamoyl chloride treated material is expressed as per cent of control values (background absorbance omitted). Dialkyl carbamoyl chloride induced cell proliferation even though no direct contact between the dialkyl carbamoyl chloride treated material and cells was obtained. Cell proliferation increased to 195% ±btcompared to 100%±1 m for untreated controls, shown in
FIG. 1 . Moreover the result shows that fibroblasts do riot bind to the dialkyl carbamoyl chloride treated material. - Twenty patients with wounds after transplantation of skin to another location are admitted to this open study. The patients are included in regularly planned follow ups with the medical staff. Inclusion criteria are the presence of a wound after skin has been removed for transplantation not more than 24 hours ago. The patients arc treated once daily using dressings manufactured according to example 3 herein. Ten of the admitted patients are treated with these cell proliferation inducing and ten patients serve as a control group treated with dressings without dialkyl carbamoyl chloride.
- At the patient's first visit to the clinic, bedside assessment of the wounds is performed. and photos or the wound areas are taken. The dressing is applied by the investigator and the patients are carefully instructed how to do the application by themselves between the controls. The patients are admitted back to the clinic at
day - The wound healing assessment is made using Bates-Jensen wound assessment tool (BWAT) as a scoring system. BWAT is described by Carrie Sussman, Barbara M. Bates-Jensen (2007); Wound care: a collaborative practice manual (third edition) Lippincott Williams & Wilkins p. 176-179. In this study we examine the cell proliferation in wounds and the following evaluation is performed through photographs by a blinded independent observer according to protocol. Clinical and visual evaluation are done bedside and through photographs according to a preset evaluation protocol. All twenty patients complete the study. No side effects are noted.
- Pictures of the lesion are taken at
day FIG. 2 . Patient treated with cell proliferation inducing dressings of Example 3 exhibits a more rapid wound healing process than the control group treated with dressings without dialkyl carbamoyl chloride. - Fibroblasts cultured according to example 1 were grown until confluence at the start of the experiment. A mechanical injury was created by scraping the monolayer of cultured cells with a 10 μl sterile plastic pipette (D10ST, Gilson, INc.), forming a cross. The resulting damage was measured with a calibrated size marker at several time points, using a image processing software. In this in vitro model, with a monolayer of cells, cell proliferation and migration was identified by measuring the distance between the cells. To accomplish this, a line was drawn parallel to the wound edges, with multiple lines then drawn perpendicular to this, across the damaged area. This allowed a mean value for the length of the damaged area to be calculated. To reproduce measurements between incubation times, calculations were always made in the same the same area. Creating a cross in each culture well made it easy to identify this area by locating the centre of the cross and always beginning in the upper right arm of the cross.
- Healing rate was calculated by dividing the restored distance by the time between each measurement. Measurements were performed at 1, 3, 6, 24, 32, 48, 56 and 72 hours, after creating the mechanical damage. In the wound-healing model, the compresses were cut into small pieces using the 5 mm biopsy punch (used in example 2). A total of 12 wells was used, with six wells used for the dialkyl carbamoyl chloride treated wound dressings as described in example 5 and six wells with medium only. Care was taken not to contaminate the culture wells with dressing debris or fibres.
- Mechanical damage to the fibroblastic surface resulted in a denuded injury averaging a distance of 530±40 μm between the edges, with no significant difference between dialkyl carbamoyl chloride treated dressings and control wells (p=0.15). In all experiments using the dialkyl carbamoyl chloride dressing (n=12), the mechanical injury was fully recovered within 72 hours, while the cell culture wells with the control medium (n=12) failed to reach complete healing in the same period. The presence of the dressing caused the fibroblasts to migrate across the damaged area more rapidly than cells in medium only. This was illustrated with photomicrographs, with a greater number of fibroblasts repairing the denuded area in cells with the dialkyl carbamoyl chloride treated dressing, compared with cells treated with only the medium (results not shown).
- For total fibroblastic restoration, a significant difference between the dialkyl carbamoyl chloride treated dressing and the untreated control medium was observed between 56 and 72 hours (
FIG. 3 ). - In this example we manufacture a standard wound dressing based on the invention in the following manner:
- The hydrophobic layer is preferably produced by applying to a cellulose acetate fabric an amount of dialkyl carbamoyl chloride, making a covalent bond between the materials. The dialkyl carbamoyl chloride is applied by letting the fabric pass through a bath of 0.5-2% carbamoyl chloride solution. The applied amount is checked by using the hydrophobicity test below.
- The acetate fabric is on rolls of 50 m length and at a width of 1 m, and taken as such to the next step.
- A fluid comprising formamide and ethylene glycol-monoethyl-ether with known dyne level between 37 and 50 dyne/cm.
- Pipettes
- Chronometer
- This control is conducted in the beginning and the in the end of every roll. The test sample of acetate woven is 0.1 m2.
- Test method:
-
Alternative 1. The impregnated woven is stretched over a frame. Using the pipette a drop of liquid with a known dyne level is placed on the woven from approximately 5 mm's height. The chronometer is started. The drop shall stay on top of the woven for at least 30 seconds and if the drop is penetrating (flowing through) the woven, the test is reperformed with higher dyne level. - Alternative 2. This alternative is used when the woven material is sparse. Pour a fluid with known dyne level into a petri dish. Remove a fiber from the woven material and place it on the surface of the fluid. The fiber shall he floating during 30 seconds and if the fiber is sinking, the test needs to be reperformed with a fluid with higher dyne level.
- Documentation. The result is documented for the batch in question.
Results. The sample shall have a dyne level less than 42 dyne/cm
Cutting and adding of carrier layer. The now bonded hydrophobic layer is cut into suitable size pieces for the final product, in thiscase 30 mm×30 mm. The cut pieces are placed on carrier-film, prepared with the adhesive and release paper and in strips of 100 mm. The bonded pieces are centered on the carrier strips and with a distance from center to center of 80 mm. A release paper of the same width is finally then applied on inner side. The now completely assembled dressing, is cut into pieces of 80 mm×100 mm, sterilized and then packed 10 by 10 in cartons.
Claims (19)
1. A method for enhancing proliferation of cells, comprising bringing cells into contact with a hydrophobic substance.
2. The method according to claim 1 , wherein said method is for use in healing of wounds.
3. The method according to claim 1 , wherein said hydrophobic substance is selected from the group consisting of dialkyl carbamoyl and alkyl ketene dimer.
4. The method according to claim 2 , wherein said hydrophobic substance is selected from the group consisting of dialkyl carbamoyl and alkyl ketene dimer and pharmaceutically acceptable salts and esters thereof.
5. The method according to claim 3 , wherein said hydrophobic substance is a halide salt of dialkyl carbamoyl, preferably dialkyl carbamoyl chloride.
6. The method according to claim 4 , wherein said hydrophobic substance is a halide salt of dialkyl carbamoyl, preferably dialkyl carbamoyl chloride.
7. A composition for use in enhancing proliferation of cells, comprising a hydrophobic substance, and optionally pharmaceutically acceptable carriers and excipients.
8. The composition according to claim 7 , wherein said composition is a dressing.
9. The composition according to claim 8 , for use in wound healing.
10. The composition according to claim 7 , wherein said hydrophobic substance is selected from dialkyl carbamoyl and alkyl ketene dimer, and pharmaceutically acceptable salts and esters thereof.
11. The composition according to claim 8 , wherein said hydrophobic substance is selected from dialkyl carbamoyl and alkyl ketene dimer, and pharmaceutically acceptable salts and esters thereof.
12. The composition according to claim 9 , wherein said hydrophobic substance is selected from dialkyl carbamoyl and alkyl ketene dimer, and pharmaceutically acceptable salts and esters thereof.
13. The composition according to claim 7 , wherein said hydrophobic substance is a halide salt of dialkyl carbamoyl, preferably dialkyl carbamoyl chloride.
14. The composition according to claim 8 , wherein said hydrophobic substance is a halide salt of dialkyl carbamoyl, preferably dialkyl carbamoyl chloride.
15. The composition according to claim 9 , wherein said hydrophobic substance s a halide salt of dialkyl carbamoyl, preferably dialkyl carbamoyl chloride.
16. The composition according to claim 10 , wherein said hydrophobic substance is a halide salt of dialkyl carbamoyl, preferably carbamoyl chloride.
17. The composition according to claim 11 , wherein said hydrophobic substance is a halide salt of dialkyl carbamoyl, preferably dialkyl carbamoyl chloride,
18. The composition according to claim 12 , wherein said hydrophobic substance is a halide salt of dialkyl carbamoyl, preferably dialkyl carbamoyl chloride.
19. The composition according to claim 7 , wherein the cells are selected from fibroblasts and keratinocytes.
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US13/623,564 US20130072565A1 (en) | 2011-09-21 | 2012-09-20 | Method of increasing cell proliferation by advanced wound dressings |
PCT/SE2012/051001 WO2013043115A1 (en) | 2011-09-21 | 2012-09-21 | Method of increasing cell proliferation by advanced wound dressings |
EP12833333.3A EP2758045A4 (en) | 2011-09-21 | 2012-09-21 | Method of increasing cell proliferation by advanced wound dressings |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201161626136P | 2011-09-21 | 2011-09-21 | |
US13/623,564 US20130072565A1 (en) | 2011-09-21 | 2012-09-20 | Method of increasing cell proliferation by advanced wound dressings |
Publications (1)
Publication Number | Publication Date |
---|---|
US20130072565A1 true US20130072565A1 (en) | 2013-03-21 |
Family
ID=47881243
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/623,564 Abandoned US20130072565A1 (en) | 2011-09-21 | 2012-09-20 | Method of increasing cell proliferation by advanced wound dressings |
Country Status (3)
Country | Link |
---|---|
US (1) | US20130072565A1 (en) |
EP (1) | EP2758045A4 (en) |
WO (1) | WO2013043115A1 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017046629A1 (en) | 2015-09-15 | 2017-03-23 | Stellenbosch University | A method of culturing cells |
US20170226456A1 (en) * | 2011-11-08 | 2017-08-10 | Dai Nippon Printing Co., Ltd. | Method for producing cell culture vessel |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB9206508D0 (en) * | 1992-03-25 | 1992-05-06 | Jevco Ltd | Biopolymer composites |
US6040493A (en) * | 1998-04-24 | 2000-03-21 | Replication Medical, Inc. | Bioreactor wound dressing |
CA2296311A1 (en) * | 1999-01-28 | 2000-07-28 | Universite Laval | Enzymatic hydrolysate of milk proteins |
US9463119B2 (en) * | 2007-01-18 | 2016-10-11 | Abigo Medical Ab | Wound dressing with a bacterial adsorbing composition and moisture holding system |
-
2012
- 2012-09-20 US US13/623,564 patent/US20130072565A1/en not_active Abandoned
- 2012-09-21 EP EP12833333.3A patent/EP2758045A4/en not_active Withdrawn
- 2012-09-21 WO PCT/SE2012/051001 patent/WO2013043115A1/en active Application Filing
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20170226456A1 (en) * | 2011-11-08 | 2017-08-10 | Dai Nippon Printing Co., Ltd. | Method for producing cell culture vessel |
WO2017046629A1 (en) | 2015-09-15 | 2017-03-23 | Stellenbosch University | A method of culturing cells |
CN108350420A (en) * | 2015-09-15 | 2018-07-31 | 斯坦陵布什大学 | The method for cultivating cell |
Also Published As
Publication number | Publication date |
---|---|
WO2013043115A1 (en) | 2013-03-28 |
EP2758045A4 (en) | 2015-05-27 |
EP2758045A1 (en) | 2014-07-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6407911B2 (en) | Hemostatic composition and therapy | |
ES2767252T3 (en) | Umbilical cord amniotic membrane products | |
ES2326873T3 (en) | SURGICAL DEVICE FOR TREATMENT OR SKIN ANALYSIS. | |
RU2308954C2 (en) | Pharmaceutical composition containing blood plasma or serum for treatment of wounds | |
Mohammadi et al. | Effect of amniotic membrane on graft take in extremity burns | |
Zajicek et al. | New biological temporary skin cover Xe-Derma® in the treatment of superficial scald burns in children | |
US9439950B2 (en) | Activated leukocyte conditioned supernatant and uses for wound healing | |
ES2467668T3 (en) | Pharmaceutical composition, bandage and method to treat skin lesion, intermediate composition and process to prepare said bandage and use of cerium salt associated with a collagen matrix | |
US20130072565A1 (en) | Method of increasing cell proliferation by advanced wound dressings | |
RU2512681C2 (en) | Chronic wound and/or wound chamber healing technique | |
RU2657806C2 (en) | Method for regional treatment of trophic ulcers | |
Newman et al. | Outbreak of atypical mycobacteria infections in US patients traveling abroad for cosmetic surgery | |
Sabetkam et al. | Impact of Mummy Substance on Proliferation and Migration of Human Wharton's Jelly-Derived Stem Cells and Fibroblasts in an In Vitro Culture System. | |
Altmeyer et al. | Wound healing and skin physiology | |
WO2024123237A1 (en) | Use of conditioned medium of cord lining mesenchymal stem cells for healing and/or preventing the formation and/or recurrence of a wound or a damaged skin area | |
JP7502803B2 (en) | Method for producing cultured tissue and topical agent | |
Bhattacharya et al. | Use of Fetal Skin and Amniotic Fluid Dressing for Non-healing Burn | |
Kreidstein | Excess skin following weight loss: cutis pleonasmus | |
CN104027358A (en) | Application of immortalized human lung microvascular endothelial cell complex cell factor in preparing biological agent for repairing ischemic tissue injury | |
Azzena et al. | Primary hypotension: a contraindication to free tissue transfer? | |
Gürlek et al. | A total lower lip reconstruction with a composite radial forearm-palmaris longus tendon flap: a clinical series | |
Bhathena | The vacuum expandable condom mold for reconstruction of the vagina | |
CN112933096A (en) | Use of butyryl timolol in preparation of medicine for treating superficial, mixed or deep hemangioma | |
Kuroyanagi | Skin Bioscience: A Molecular Approach | |
Narayan | Vascular lesions of the lip: an aid to resection |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |